Abstract
Over the years fluorescence correlation spectroscopy (FCS) has proven to be a useful technique that has been utilized in several fields of study. Although FCS initially suffered from poor signal-to-noise ratios, the incorporation of confocal microscopy has overcome this drawback and transformed FCS into a sensitive technique with high figures of merit. In addition, tandem methods have evolved to include dual-color cross-correlation, total internal reflection fluorescence correlation, and fluorescence lifetime correlation spectroscopy combined with time-correlated single-photon counting. In this review, we discuss several applications of FSC for biochemical, microfluidic, and cellular investigations.
🔬 Techniques
✨ Fluorophores
🧪 Sample Preparation
🔬 Cell Lines
🏭 Microscope Brands
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1
Conceptual diagram of a fluctuating fluorescence signal (A) as a function of time. The fluctuation of the signal is used to calculate the autocorrelation, G(u03c4), where u03c4 is the lag time from th...
Figure 2
A typical confocal FCS system. Laser light is focused by an objective (usually with high numerical aperture) to a diffraction limited spot. Fluorescence is collected by the same objective and filtered...
Figure 3
FCS measurements of flow in a low-shear cell culture chip. Laminar flow is disrupted at a T-channel interface, and FCS was used to probe the flow velocity in the junction. Flow was slowest in the cent...
Figure 4
(A) Graphic visualization of the 2fFCS confocal volume and the nomenclature we use to describe its orientation relative to the direction of flow. (B) Typical fitted 2fFCS correlation curves. The four ...
Figure 5
TIR-FCS. An evanescent wave generated using a prism or waveguide the top surface of the sample. A confocal microscope collects fluorescence from one spot, allowing signal to be restricted to close to ...
Figure 6
FCS measurements of surface receptor density. By focusing the laser spot of the cell membrane, the number of fluorophores in the probe volume (i.e. the membrane surface) can be estimated. The ratio of...
Figure 7
Schematic of soluble ligand binding on a planar membrane (a). K a and K d (association and dissociation rate constants) were later determined for IgG interacting with the planar membrane. The correlat...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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